Scanning probe microscopy elucidates gelation and rejuvenation of biomolecular condensates DOI Creative Commons
Aida Naghilou, Oskar Armbruster, Alireza Mashaghi

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Aug. 29, 2024

Abstract Comprehensive understanding of dynamics and disease-associated solidification biomolecular condensates is closely tied to analysis their mechanical characteristics. Despite recent technical advances in rheological studies condensates, these still vastly rely on methods restricted small forces, rendering measurements droplets with higher elasticities after transition solid challenging. Here, we develop assays for in-depth characterization by scanning probe microscopy. We demonstrate this technique measuring the behavior heterotypic poly-L-lysine heparin showcasing multi-route liquid gel transition, as well rejuvenation chemical alterations medium. Due wide-spread application microscopy biological fields, its capability rapid, high throughput, force range studies, integration nanoscale morphological measurements, our probe-based method a significant breakthrough investigating condensate behavior, leading accelerated development therapies.

Language: Английский

Peptide-Mediated Liquid-Liquid Phase Separation and Biomolecular Condensates DOI

Guangle Li,

Chengqian Yuan, Xuehai Yan

et al.

Soft Matter, Journal Year: 2025, Volume and Issue: 21(10), P. 1781 - 1812

Published: Jan. 1, 2025

Peptide-mediated liquid–liquid phase separation (LLPS) underpins the formation of dynamic biomolecular condensates, regulated by diverse molecular interactions, and highlights potential applications in drug delivery synthetic biology.

Language: Английский

Citations

2
Structural insights and milestones in TDP-43 research: A comprehensive review of its pathological and therapeutic advances DOI Creative Commons
Mei Dang, Longjiang Wu, Xiaoying Zhang

et al.

International Journal of Biological Macromolecules, Journal Year: 2025, Volume and Issue: unknown, P. 141677 - 141677

Published: March 1, 2025

Transactive response (TAR) DNA-binding protein 43 (TDP-43) is a critical RNA/DNA-binding involved in various cellular processes, including RNA splicing, transcription regulation, and stability. Mislocalization aggregation of TDP-43 the cytoplasm are key features pathogenesis several neurodegenerative diseases, amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), Alzheimer's disease (AD). This review provides comprehensive retrospective prospective analysis research, highlighting structural insights, significant milestones, evolving understanding its physiological pathological functions. We delineate five major stages from initial discovery as hallmark neurodegeneration to recent advances liquid-liquid phase separation (LLPS) behavior interactions with processes. Furthermore, we assess therapeutic strategies targeting pathology, categorizing approaches into direct indirect interventions, alongside modulating aberrant LLPS. propose that future research will focus on three areas: polymorphisms for disease-specific therapeutics, exploring dual temporal-spatial modulation TDP-43, advancing nano-therapy. More importantly, emphasize importance TDP-43's functional repertoire at mesoscale, which bridges molecular functions broader offers foundational framework development.

Language: Английский

Citations

2
Scanning probe microscopy elucidates gelation and rejuvenation of biomolecular condensates DOI Creative Commons
Aida Naghilou, Oskar Armbruster, Alireza Mashaghi

et al.

Cell Reports Physical Science, Journal Year: 2025, Volume and Issue: unknown, P. 102430 - 102430

Published: Feb. 1, 2025

Language: Английский

Citations

1
Making sense of blobs, whorls, and shades: methods for label-free, inverse imaging in bright-field optical microscopy DOI
Braulio Gutiérrez–Medina

Biophysical Reviews, Journal Year: 2025, Volume and Issue: unknown

Published: March 18, 2025

Language: Английский

Citations

1
Live-cell quantification reveals viscoelastic regulation of synapsin condensates by α-synuclein DOI Creative Commons
Huan Wang, Christian Hoffmann, Johannes Vincent Tromm

et al.

Science Advances, Journal Year: 2025, Volume and Issue: 11(16)

Published: April 18, 2025

Synapsin and α-synuclein represent a growing list of condensate-forming proteins where the material states condensates are directly linked to cellular functions (e.g., neurotransmission) pathology neurodegeneration). However, quantifying condensate properties in living systems has been substantial challenge. Here, we develop micropipette aspiration whole-cell patch-clamp (MAPAC), platform that allows direct quantification live cells. We find 10,000-fold variations viscoelasticity synapsin condensates, regulated by partitioning α-synuclein, marker for synucleinopathies. Through vitro reconstitutions, identify multiple molecular factors distinctly regulate viscosity, interfacial tension, maturation confirming roles α-synuclein. Overall, our study provides unprecedented quantitative insights into neuronal reveals crucial role regulating viscoelasticity. Furthermore, envision MAPAC applicable broad range vivo.

Language: Английский

Citations

1
From superior contrast to super resolution label free optical microscopy DOI Creative Commons
Nikhil Jayakumar, Balpreet Singh Ahluwalia

npj Imaging, Journal Year: 2025, Volume and Issue: 3(1)

Published: Jan. 13, 2025

Abstract Label-free optical microscopy utilizes the information encoded in light scattered off unlabeled particles to generate images. This review article starts with a discussion on how this matter interaction gives rise issues of poor-contrast and diffraction-limited spatial resolution. Then, reviews various far-field label-free techniques that have been developed, an emphasis physical mechanisms behind image formation processes such techniques. Thus aims elucidate state-of-the-art their current applications.

Language: Английский

Citations

0
AFM multiparametric characterization of LLPS in plants DOI
Junbao Pu, Shanshan Wu, Liang Chen

et al.

Trends in Plant Science, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

Language: Английский

Citations

0
ExPOSE: a comprehensive toolkit to perform expansion microscopy in plant protoplast systems DOI Creative Commons
Kevin L. Cox, Sarah A. Pardi, Lily O’Connor

et al.

The Plant Journal, Journal Year: 2025, Volume and Issue: 121(5)

Published: March 1, 2025

Expansion microscopy (ExM) achieves nanoscale imaging by physical expansion of fixed biological tissues embedded in a swellable hydrogel, enhancing the resolution any optical microscope several-fold. While ExM is commonly used animal cells and tissues, there are few plant-specific protocols. Protoplasts widely cell system across plant species, especially studying biomolecule localization. Here, we present an approach to achieve robust protoplasts, termed PrOtoplast SystEms (ExPOSE). We demonstrate that coupling ExPOSE with other techniques, immunofluorescence situ hybridization chain reaction visualize proteins mRNAs, respectively, greatly enhances spatial endogenous biomolecules. Additionally, this study, tested effectiveness versatility technique observe biomolecular condensates Arabidopsis protoplasts transcription factors maize at increased resolution. can be relatively inexpensive, fast, simple implement.

Language: Английский

Citations

0
ExPOSE: A comprehensive toolkit to perform expansion microscopy in plant protoplast systems DOI
Kevin L. Cox, Sarah A. Pardi, Lily O’Connor

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: July 16, 2024

Abstract Expansion microscopy (ExM) achieves nanoscale imaging by physical expansion of fixed biological tissues embedded in a swellable hydrogel, enhancing the resolution any optical microscope several-fold. While ExM is commonly used animal cells and tissues, there are few plant specific protocols. Protoplasts widely cell system across species, especially studying biomolecule localization. Here, we present an approach to achieve robust protoplasts, termed Ex pansion P r O toplast S yst E ms (ExPOSE). We demonstrate that coupling ExPOSE with other techniques, immunofluorescence situ hybridization chain reaction visualize proteins mRNAs, respectively, greatly enhances spatial endogenous biomolecules. Additionally, this study, tested effectiveness versatility technique observe biomolecular condensates Arabidopsis protoplasts transcription factors maize at increased resolution. can be relatively inexpensive, fast, simple implement.

Language: Английский

Citations

0
Mechanistic Insights into the c-MYC G-Quadruplex and Berberine Binding inside an Aqueous Two-Phase System Mimicking Biomolecular Condensates DOI
Susmita Pradhan,

Marco Campanile,

Shubhangi Sharma

et al.

The Journal of Physical Chemistry Letters, Journal Year: 2024, Volume and Issue: 15(34), P. 8706 - 8714

Published: Aug. 19, 2024

We investigated the binding between c-MYC G-quadruplex (GQ) and berberine chloride (BCl) in an aqueous two-phase system (ATPS) with 12.3 wt % polyethylene glycol 5.6 dextran, mimicking highly crowded intracellular biomolecular condensates formed via liquid–liquid phase separation. found that ATPS, complex formation is significantly altered, leading to increase affinity a change stoichiometry of respect neat buffer conditions. Thermodynamic studies reveal becomes more thermodynamically favorable ATPS due entropic effects, as strong excluded volume effect inside droplets reduces penalty associated binding. Finally, BCl for GQ higher than those other DNA structures, indicating potential specific interactions. Overall, these findings will be helpful design drugs targeting structures cancer-related biocondensates.

Language: Английский

Citations

0